KR20100066548A - Solid forms of (s)-2-amino-3-(4-(2-amino-6-((r)-2,2,2-trifluoro-1-(3'-methoxybiphenyl-4-yl)ethoxy)pyrimidin-4-yl)phenyl)propanoic acid and methods of their use - Google Patents

Abstract

Solid forms of (S)-2-amino-3-(4-(2-amino-6-((R)-2,2,2-trifluoro-1-(3'-methoxybiphenyl-4-yl)ethoxy)pyrimidin-4-yl)phenyl)propanoic acid and salts thereof are disclosed.

Description

(S) -2-amino-3 (4)-(2-amino-6)-((R) -2,2,2-trifluoro-1- (3'-methoxybiphenyl-4-yl)) Solid form of ethoxy) pyrimidin-4-yl) phenyl) propanoic acid and its use {SOLID FORMS OF (S) -2-AMINO-3- (4- (2-AMINO-6-((R)- 2,2,2-TRIFLUORO-1- (3'-METHOXYBIPHENYL-4-YL) ETHOXY) PYRIMIDIN-4-YL) PHENYL) PROPANOIC ACID AND METHODS OF THEIR USE}

This application claims priority to US Provisional Application No. 60 / 978,303, filed October 8, 2007, which is incorporated herein by reference in its entirety.

The present invention provides (S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3'-methoxybiphenyl-4) -Yl) ethoxy) pyrimidin-4-yl) phenyl) propanoic acid and salts thereof.

Different solid forms of the same compound may have substantially different properties. For example, the amorphous form of a drug may exhibit different dissolution properties and different bioavailability patterns than its crystalline form (s), which properties may affect how the drug should be administered to achieve optimal effects. Can be. In addition, the amorphous and crystalline forms of the drug may have different handling properties (eg, flowability, compressibility), dissolution rate, solubility and stability, all of which may affect the preparation of the dosage form. Thus, access to various forms of drugs is desirable for a variety of reasons. In addition, regulatory agencies (eg, the US Food and Drug Administration) may require the identification of all its solid forms prior to approving a product containing a new drug substance. Goho, Science News 166 (8): 122-123 (2004)].

The compounds may exist in one or more crystalline forms, but cannot be predicted with any certainty regarding the existence and properties of these forms. In addition, no standard procedure exists for preparing all possible polymorphic forms of the compound. Also, even after one polymorph has been identified, other forms of existence and properties can only be determined by further experimentation. Id.

Part of the invention is a tryptophan hydroxylase inhibitor (S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3) '-Methoxybiphenyl-4-yl) ethoxy) pyrimidin-4-yl) phenyl) propanoic acid and pharmaceutically acceptable salts thereof. Certain solid forms are crystals.

One embodiment of the present invention includes a pharmaceutical composition comprising the solid form described herein.

Some of the invention relates to (S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3'-methoxybiphenyl) -4-yl) ethoxy) pyrimidin-4-yl) phenyl) propanoic acid and pharmaceutically acceptable salts thereof. This compound is an inhibitor of tryptophan hydroxylase. When administered to animals, this compound lowers peripheral serotonin levels, which can be used to treat a wide range of diseases and disorders. See U.S. Patent Application No. 11 / 638,677, filed December 12, 2006, and 60 / 946,246, filed June 26, 2007.

The invention also provides (S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3'-methoxybiphenyl-). A dosage form comprising a solid form of 4-yl) ethoxy) pyrimidin-4-yl) phenyl) propanoic acid and a method of using the same.

5.1. Justice

Unless stated otherwise, the phrases "diseases or disorders mediated by peripheral serotonin" and "diseases and disorders mediated by peripheral serotonin" refer to diseases and conditions with one or more signs of severity affected by peripheral serotonin levels and / or Or disability.

Unless stated otherwise, the terms "manage", "managing" and "management" prevent a recurrence of such a disease or disorder in a patient who already has a particular disease or disorder, and / or have a disease or disorder Prolonging the time the patient is in a recovery state. The term includes adjusting the threshold, development and / or persistence of a disease or disorder, or changing the way a patient responds to a disease or disorder.

Unless stated otherwise, the terms "prevent", "prevention" and "prevention" take into account the actions that a patient takes before suffering from a particular disease or disorder, which inhibits or reduces the severity of the disease or disorder. . In other words, the term includes prevention.

Unless stated otherwise, a “prophylactically effective amount” of a compound is an amount sufficient to prevent or prevent recurrence of the disease or condition or one or more signs associated with the disease or condition. A prophylactically effective amount of a compound means an amount of therapeutic agent alone, or an amount of therapeutic agent in combination with other agents that provides prophylactic benefit in the prevention of a disease or condition. The term “prophylactically effective amount” may include an amount that overall improves prophylaxis or enhances the prophylactic efficacy of other prophylactic agents.

Unless stated otherwise, a “therapeutically effective amount” of a compound is an amount sufficient to provide therapeutic benefit in the treatment or management of the disease or condition, or to delay or minimize one or more signs associated with the disease or condition. A therapeutically effective amount of a compound means an amount of therapeutic agent alone, or an amount of therapeutic agent in combination with other therapies that provide therapeutic benefit in the treatment or management of the disease or condition. The term “therapeutically effective amount” may include an amount that overall improves the therapy, reduces or avoids the signs or causes of the disease or condition, or improves the therapeutic efficacy of other therapeutic agents.

Unless stated otherwise, the terms “treat”, “treating” and “treatment” take into account actions taken while a patient is suffering from a particular disease or disorder and may reduce the severity of the disease or disorder or one or more of its symptoms. Or to delay or slow the progression of a disease or disorder.

Unless stated otherwise, the term "comprises" has the same meaning as "including, but not limited to". Similarly, the term "such as" has the same meaning as the term "but not limited to, for example".

Unless stated otherwise, it is understood that one or more adjectives immediately preceding the set of nouns applies to each noun. For example, the phrase "optionally substituted alkyl, aryl, or heteroaryl" has the same meaning as "optionally substituted alkyl, optionally substituted aryl, or optionally substituted heteroaryl".

It is also noted that any atom depicted as having unsatisfied valences in the figures is attached with sufficient hydrogen atoms to assume that the valences are satisfied. In addition, chemical bonds represented by one solid line parallel to one dashed line include both single and double (eg aromatic) bonds, if valences are allowed. While not displaying stereochemistry (eg, in bold or dotted lines), structures showing compounds having one or more chiral centers include pure stereoisomers and mixtures thereof (eg racemic mixtures). Similarly, the names of compounds having one or more chiral centers, although not specifying the stereochemistry of their centers, include pure stereoisomers and mixtures thereof.

5.2. Solid form

The present invention provides (S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3'-methoxybiphenyl-4) -Yl) ethoxy) pyrimidin-4-yl) phenyl) propanoic acid:

And solid forms of their salts. Specific salts are crystals. Specific salts include tosylate and maleate salts.

One embodiment of the invention provides (S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3'-methoxy) Anhydride and hydrated crystalline forms of biphenyl-4-yl) ethoxy) pyrimidin-4-yl) phenyl) propanoate tosylate.

Certain forms of this compound have a (starting temperature) (S) -2-amino-3- (4- (2-amino-6-((R) -2,2) having a melting point of about 241 ° C. as measured by DSC. , 2-trifluoro-1- (3'-methoxybiphenyl-4-yl) ethoxy) pyrimidin-4-yl) phenyl) propanoate tosylate anhydride. In this regard, the term "about" means ± 5.0 ° C. The form provides an X-ray diffraction (XRPD) pattern containing peaks at one or more of the 2θ angles of about 3.5, 7.0, 8.6, 10.9, 13.5, 14.0, 15.1, 17.3, and / or 20.5. In this regard, the term "about" means ± 0.3 degrees. As will be appreciated by those skilled in the art, the relative intensity of the peaks in the XRPD pattern of the crystalline material may vary depending on how the sample was prepared and how the data was collected. With this in mind, an example of the XRPD pattern of the crystalline form is provided in FIG. 1.

Another form of this compound has a melting point of about 221 ° C. as measured by DSC (initiation of endothermic peak with a maximum at about 227 ° C.) (S) -2-amino-3- (4- (2-amino- 6-((R) -2,2,2-trifluoro-1- (3'-methoxybiphenyl-4-yl) ethoxy) pyrimidin-4-yl) phenyl) propanoate tosylate 1 It is a hydrate. In this regard, the term "about" means ± 5.0 ° C. The form provides an XRPD pattern containing peaks at one or more of 2θ angles of about 3.6, 8.2, 8.7, 13.1, 14.5, 17.5, 18.0, 19.9 and / or 21.4. In this regard, the term "about" means ± 0.3 degrees. An example of the XRPD pattern of the crystalline form is provided in FIG. 2. 3 provides an example of a Raman spectrum of this type.

Another form of this compound is a (S) -2-amino-3- (4- (2-amino) having a melting point of about 238 ° C. (initiation of an endothermic peak having a maximum at about 242 ° C.) as measured by DSC. -6-((R) -2,2,2-trifluoro-1- (3'-methoxybiphenyl-4-yl) ethoxy) pyrimidin-4-yl) phenyl) propanoate tosylate It is a dihydrate. In this regard, the term "about" means ± 5.0 ° C. The form provides an XRPD pattern containing peaks at one or more of the 2θ angles of about 8.6, 9.0, 17.2, 17.8, 18.6, 21.6, 25.2 and / or 26.9. In this regard, the term "about" means ± 0.3 degrees. An example of the XRPD pattern of the above type is provided in FIG. 4.

Another embodiment of the invention is (S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3'-meth). Anhydride and hydrated forms of oxybiphenyl-4-yl) ethoxy) pyrimidin-4-yl) phenyl) propanoate maleate.

The present invention includes solids that are mixtures of both amorphous and crystalline forms. Such specific solids are crystals (S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3'-methoxybiphenyl) -4-yl) ethoxy) pyrimidin-4-yl) phenyl) propanoic acid or a pharmaceutical salt thereof in an amount of about 50, 75, 80, 85, 90, 95 or 99% by weight or more.

(S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3'-methoxybiphenyl-4-yl) Crystalline salts of ethoxy) pyrimidin-4-yl) phenyl) propanoic acid can be prepared by heating a solution comprising the compound and a pharmaceutically acceptable acid, reducing the solubility of the resulting salt, and isolating the crystalline salt. . In one embodiment, the solution is in THF / water. In certain methods, the THF / water solution is heated to about 40 ° C to 60 ° C. Crystallization of the salt is then caused by adding anti-solvent (eg acetonitrile) to the hot solution and then cooling it.

5.3. How to treat

The present invention includes a method of inhibiting tryptophan hydroxylase (TPH), which method comprises contacting TPH with a compound of the invention (ie, a compound disclosed herein). In certain methods, TPH is the TPH1 isotype. In another method, TPH is the TPH2 isotype. In certain methods, inhibition is in vitro. In another method, the inhibition is in vivo.

The present invention encompasses methods of treating, preventing and managing various diseases and disorders mediated by peripheral serotonin, which methods include inhibiting TPH1 activity in patients in need of such treatment, prevention or management.

Certain diseases and disorders include carcinoid syndrome and gastrointestinal diseases and disorders. Examples of specific diseases and disorders include abdominal pain (eg, associated with medullary carcinoma of the thyroid gland), anxiety, carcinoid syndrome, celiac disease, constipation (eg, constipation with idiopathic causes and idiopathic Constipation), Crohn's disease, depression, diabetes, diarrhea (eg bile acid diarrhea, enterotoxin-induced secretory diarrhea, diarrhea with a cause of cause, idiopathic diarrhea (eg idiopathic secretory diarrhea), and traveler's diarrhea ), Vomiting, functional abdominal pain, functional dyspepsia, irritable bowel syndrome (IBS), lactose intolerance, MEN types I and II, Ogilvie's syndrome, Pancreatic Cholera Syndrome, pancreatic insufficiency, chromaffin cell tumor , Scleroderma, somatization disorder, and Zolinger-Ellison syndrome.

In certain methods of the invention, the treatment, management and / or prevention of a disease or disorder is accomplished while avoiding side effects associated with changes in central nervous system (CNS) serotonin levels. Examples of such side effects include anxiety, anxiety disorders, depression, and sleep disorders (eg, insomnia and sleep arousal disorders).

5.4. Pharmaceutical composition

The present invention includes pharmaceutical compositions and dosage forms comprising the solid forms of the invention. Pharmaceutical compositions and dosage forms of the invention may optionally contain one or more pharmaceutically acceptable carriers or excipients. Certain pharmaceutical compositions may be administered orally, topically, to mucous membranes (eg, nasal, pulmonary, sublingual, vaginal, buccal, or rectal), parenteral (eg, subcutaneous, intravenous, bolus injection, muscle) Single unit dosage forms suitable for intra-, or intra-arterial, or transdermal administration. Examples of dosage forms include tablets; Dragees; Capsules such as soft elastic gelatin capsules; Kake; Trocheses; Lozenges; Dispersants; Suppositories; Ointment; Poultices (foam agents); Paste; Powder; dressing; cream; Plaster; Solution; patch; Aerosols (eg nasal sprays or inhalants); Gels; Liquid dosage forms including suspensions (eg, aqueous or non-aqueous liquid suspensions, oil-in-water emulsions, or water-in-oil liquid emulsions), solutions, and elixirs, suitable for oral or mucosal administration to a patient; Liquid dosage forms suitable for parenteral administration to a patient; And sterile solids (eg, crystalline or amorphous solids) that can be reconstituted to provide liquid dosage forms suitable for parenteral administration to a patient.

The formulation should suit the mode of administration. For example, oral administration may require an enteric coating to protect the active ingredient from degradation in the gastrointestinal tract. In another embodiment, the active ingredient may be administered in a liposome formulation to block it from the degrading enzymes, facilitate transport in the circulation, and / or deliver across the cell membrane to intracellular regions.

The composition, shape and type of dosage forms of the invention will typically vary depending on their use. For example, a dosage form used for the acute treatment of a disease may contain more of one or more active ingredients it contains compared to dosage forms used for chronic treatment of the same disease. Similarly, parenteral dosage forms may contain less than one or more active ingredient that they comprise as compared to oral dosage forms used to treat the same disease. It will be readily appreciated by those skilled in the art that in these and other ways the specific dosage forms encompassed by the present invention will vary from one another. See, eg, Remington's Pharmaceutical Sciences, 18 ^th ed., Mack Publishing, Easton PA (1990).

Certain aspects of the present invention will be understood with reference to the accompanying drawings.
1 shows anhydride (S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3'-methoxybiphenyl-) X-ray diffraction pattern of the crystalline solid form of 4-yl) ethoxy) pyrimidin-4-yl) phenyl) propanoic acid tosylate. This diffractogram was obtained using a Rigaku MiniFlex diffractometer (Cu Kα radiation).
Figure 2 shows (S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3'-methoxybiphenyl-4) X-ray diffraction pattern of the crystalline solid form of -yl) ethoxy) pyrimidin-4-yl) phenyl) propanoic acid tosylate monohydrate. This diffractogram was obtained using a Bruker D8 Advance diffractometer (Cu Kα radiation).
Figure 3 shows (S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3'-methoxybiphenyl-4) FT-Raman spectrum of the crystalline solid form of -yl) ethoxy) pyrimidin-4-yl) phenyl) propanoic acid tosylate monohydrate. This spectrum was obtained using a Bruker RFS100 spectrometer (1064 nm excitation).
Figure 4 shows (S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3'-methoxybiphenyl-4) X-ray diffraction pattern of the crystalline solid form of -yl) ethoxy) pyrimidin-4-yl) phenyl) propanoic acid tosylate dihydrate. This diffractogram was obtained using a Rigaku miniplex diffractometer (Cu Kα radiation).

6.1. Preparation of (R) -1- (4-bromophenyl) -2,2,2-trifluoroethanol

The compounds are described in Okhuma, et al. J. Am. Chem. Soc., 120: 13529-13530 (1998). 4-bromo-trifluoroacetophenone ( 1 , Wilmington PharmaTech, Delaware, USA, 100.0 g, 395 mmol), potassium tert-butoxide (2-methyl-) in a 1 L high pressure vessel 1 M solution in 2-propanol, 5.0 ml, 10.0 mmol, 0.025 eq), and catalyst [(trans) -RuCl ₂ [(R) -Xyl-P-Phos] [(R) -DIAPEN] (New Jersey, USA) Johnson Matthey, 200 mg, 0.16 mmol, 0.04% mol) was charged. This mixture was dissolved in anhydrous 2-propanol (175 ml) and the whole vessel was purged with argon in 3 vacuum-thaw cycles. The reaction mixture was then purged with hydrogen in three vacuum-thaw cycles. The reaction was performed under 60 psi hydrogen atmosphere. If no more hydrogen was consumed after stirring for 24 hours, the reaction was considered complete by GC-MS analysis (the starting material no longer exists ketone). The contents of the reaction vessel were transferred to a round bottom flask, washed with MeOH (3 × 20 ml), and concentrated under reduced pressure until no solvent was distilled off. The resulting orange-brown oil was then dissolved in heptane (1000 ml), washed with water (2 × 100 ml) and brine (100 ml) and dried over sodium sulfate. To the dried organic layer was added Darco® activated carbon (20 g) and Hyflo® supercell (20 g) and the mixture was heated at 70 ° C. for 1 hour. The mixture was filtered hot to give a pale yellow solution. The filtrate was concentrated under reduced pressure with heating (about 50 ° C. to 60 ° C.) until no more solvent was distilling off. The resulting yellow oil was dissolved in 60 ° C. warmed heptane (350 ml), stirred and cooled. As the temperature cooled to room temperature, a white solid began to precipitate. After stirring for 4 hours, the solid was filtered and dried to give the title product as a white powder (63.5 g, 63%,> 99% ee).

6.2. Preparation of (S) -1- (4-bromophenyl) -2,2,2-trifluoroethanol

Using a process similar to that in the above examples, using a catalyst [(trans) -RuCl ₂ [(S) -Xyl-P-Phos] [(S) -DIAPEN] (Johnson Matty, NJ) The compound was prepared.

6.3. Preparation of (R) -2,2,2-trifluoro-1- (p-tolyl) ethanol

Similarly, 2,2,2-trifluoro-1- (p-tolyl) ethane using catalyst [(trans) -RuCl ₂ [(R) -Xyl-P-Phos] [(R) -DIAPEN] Hydrogenation of the one gave the title compound.

6.4. Preparation of (S) -2,2,2-trifluoro-1- (p-tolyl) ethanol

Similarly, the title compound was prepared using catalyst [(trans) -RuCl ₂ [(S) -Xyl-P-Phos] [(S) -DIAPEN].

6.5. Preparation of (R) -2,2,2-trifluoro-1- (3'-methoxybiphenyl-4-yl) ethanol

(R) -1- (4-Bromophenyl) -2,2,2-trifluoroethanol ( 2 , 69 g, 0.27 mol,> 99% ee) in ethanol (560 ml), 3-methoxy phenyl Boronic acid (Matrix, 51 g, 0.34 mol, purity 97%), and bis (triphenylphosphine) palladium (II) dichloride (0.95 g, 0.5% mol) in a stirred solution in water (140 ml) A solution of potassium carbonate (112 g, 0.81 mol) was added under nitrogen. The resulting mixture was heated at 75 ° C. for 1 h and considered complete by GC-MS or TLC. After the reaction mixture was cooled to 40 ° C., it was filtered through a pad of Celite and washed with methanol (3 × 100 ml). The filtrate was diluted with 100 ml of water and concentrated. The resulting syrup was dissolved in 700 ml of ethyl acetate and washed with 1N sodium hydroxide (2 × 100 ml), water (2 × 100 ml) and brine (1 × 100 ml). The organic layer was heated with activated carbon (14 g) and Hyflo super cell (14 g) at 60 ° C. for 1 hour. The mixture was filtered hot, washed with ethyl acetate (100 ml) and concentrated to syrup. This syrup was immediately dissolved in 1% ethyl acetate / heptane (700 ml) and stirred for 4 hours. The resulting slurry was filtered and dried to give the title compound as a white crystalline solid ( 3 , 68 g, 89% yield,> 99% ee).

In an alternative crystallization method, the crude product syrup / solid (10 g) was dissolved in MTBE (10 ml) and diluted with heptane (200 ml). This solution was concentrated to about 70 ml under reduced pressure. The mixture was stirred at rt overnight and the resulting slurry was filtered and dried to give the title compound ( 3 , 8.8 g) as a white crystalline solid.

6.6. Preparation of (R) -2,2,2-trifluoro-1- (3'-methoxybiphenyl-4-yl) ethanol

A 22 L round bottom flask equipped with a mechanical stirrer, thermocouple attached to a temperature control, and a condenser with nitrogen line was charged with compound 2 (1.00 kg, 1 wt, 3.92 mol) and ethanol (4.5 L, 4.5 vol). . The mixture was sparged with nitrogen for 10 minutes and (Ph ₃ P) ₂ PdCl ₂ (12.6 g, 0.0126 wt, Strem) was added. Nitrogen was further sparged and a solution of K ₂ CO ₃ (1.63 kg, 3 equiv) in water (2 vol) was added. The mixture was heated to 75 ° C. under nitrogen and then a solution of approximately 20% 3-methoxy phenylboronic acid (715 g, 4.70 mol, 1.2 equiv, Usun) in ethanol (4.5 vol) was added via peristaltic pump. It was. After 20 minutes, in-process control (IPC) samples were taken and showed that boronic acid was consumed. This process was repeated until all boronic acid was added. HPLC analysis after stirring for an additional 20 minutes indicated the reaction was complete. Heat was blocked and water (3.6 vol) was added at 69 ° C. The reaction mixture was then filtered through a pad of Celite (Celpure P300, 0.15 wt., Sigma) at 50 ° C. and the filter cake washed with methanol (2 × 2.5 vol). The filtrate was concentrated to 5 vol at 40 ° C. to 45 ° C. under reduced pressure. The slurry was then transferred to a separatory funnel and MTBE (10 vol) was added. The mixture was then washed with 50% sodium hydroxide solution (0.6 vol). After stirring, the layers were separated and the aqueous phase was extracted with MTBE (1.5 vol). The organic extracts were combined and washed with water (1 vol) followed by 20% aqueous sodium chloride (1 vol) to give 11.9 volumes of organic product solution. This solution was transferred to the reactor and treated with a Dalco G-60 (0.3 wt) slurry in MTBE (1 vol) and heated to 50 ° C. After 90 minutes, the mixture was filtered through a CellPure P300 pad (0.15 wt) and washed with MTBE (2 × 3 vol).

The filtrate (14.8 vol) was transferred to the reactor and distilled under vacuum at 45 ° C. to remove MTBE. The filtrate was reduced to 6.7 volumes over 2.5 hours, then heptane (3.15 vol) was added. The solution was further distilled to 6.7 vol over 1 h at 50 ° C., then additional heptane (3.15 vol) was added. The solution was concentrated to 6.7 vol at 55 ° C. over 1.5 h and then heptane was added (3.15 vol). Precipitation was immediately observed and distillation continued at 60 ° C. under vacuum. After 2.5 hours the distillation was stopped (7 vol remaining), the heat was cut off and the batch was cooled to ambient temperature overnight. The batch was filtered at 24 ° C. and washed with heptane (1.5 vol). The solid was dried over vacuum at rt under vacuum to give 799.7 g of 3 as a white solid [yield 72%,> 99% (AUC)].

6.7. Preparation of (R) -2,2,2-trifluoro-1- (3'-fluorobiphenyl-4-yl) ethanol

Similar to the above procedure, the title compound was prepared from (R) -1- (4-bromophenyl) -2,2,2-trifluoroethanol ( 2 ) and 3-fluorophenylboronic acid.

6.8. Preparation of (S) -methyl 2- (tert-butoxycarbonylamino) -3- (4- (trifluoromethylsulfonyloxy) phenyl) propanoate

This compound is described in Shiehh, et al. J. Org. Chem., 1992, 57, 379-381]. Triple anhydride (in a solution of Boc-Tyr-OMe ( 4 , Bachem, CA, 100 g, 0.34 mol) and N-methylmorpholine (51 g, 1.5 eq) in dichloromethane (1000 ml) 100 g, 1.05 eq) was added at -5 ° C to -15 ° C over 2 hours. The resulting red solution was stirred at -10 ° C for 10 minutes. HPLC analysis showed that the starting material was completely lost. The reaction was quenched with 10% citric acid (500 ml). The organic layer was washed with 10% citric acid (500 ml) followed by water (500 ml). The resulting pale pink solution was concentrated to 200 ml under reduced pressure. It was diluted with acetonitrile (600 ml) and further concentrated to 200 g of solution. This solution was used in the next step without further purification. The expected yield was 98% by stripping the sample to dryness to obtain a low melting pale yellow solid.

6.9. (S) -2- (Tert-butoxycarbonylamino) -3- (4- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) phenyl) Preparation of Propanoic Acid

Ester compound 6 was prepared based on the procedure in Firooznia, et al., Tetrahedron Lett., 1999, 40, 213-216. Bis (pinacolato) diboron (90 g, 1.1 eq), potassium acetate (63 g, 2 eq), tricyclohexylphosphine (2.3 g, 2.5% mol), and palladium acetate (0.72 g, 1 mol% ) Was mixed in acetonitrile (950 ml) and the resulting mixture was stirred at room temperature for 5 minutes. The triflate ( 5 ) solution (190 g, 0.32 mol) was added and the resulting mixture was heated and cooled at 80 ° C. for 1 hour. HPLC showed that the starting material was consumed completely. The reaction mixture was quenched with aqueous potassium bicarbonate solution (57 g in 475 ml of water) and the resulting mixture was stirred at room temperature for 30 minutes. The mixture was filtered through a 20 μl cellulose pad to remove palladium black. The organic layer sample was concentrated and purified by column chromatography (gradient: 1:10 to 1: 4 ethyl acetate / hexanes) to give ester compound 6 as a clear oil.

The organic layer of 6 was stirred with aqueous lithium hydroxide solution (23 g in 500 ml of water) at room temperature for 30 minutes. The pH of the resulting slurry was adjusted to about 10 with 6 N hydrochloric acid and filtered. The cake was washed with water (200 ml). Acetonitrile was removed from the filtrate under reduced pressure to give an aqueous slurry (950 ml, additional water was added during distillation). The slurry was filtered through a pad of 20 μm cellulose and washed with water (200 ml). The filtrate was washed with MTBE (500 ml) and diluted again with 700 ml MTBE. The mixture was acidified to pH about 4.5 with 6 N hydrochloric acid. The organic layer was washed with water (500 ml) and concentrated under reduced pressure to give the title product ( 7 ) as a brown oil (206 g, yield 95% based on purity expected by NMR). The crude product was used directly in the next step.

Compound 7 was also isolated by crystallization. For example, the MTBE solution of 7 was dried over anhydrous Na ₂ SO ₄ and concentrated to about 1.0 vol under vacuum. Heptane (2.5 vol) was added and concentrated to about 1.5 vol under vacuum. Heptane (4.2 vol) was added slowly at 36 ° C. to 42 ° C. and then slowly cooled to 5 ° C. to 10 ° C. The resulting slurry was filtered, washed with heptane and dried at 20 ° C. to 30 ° C. under vacuum to give product 7 in about 76% yield.

6.10. (S) -2- (Tert-butoxycarbonylamino) -3- (4- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) phenyl) Alternative crystallization of propanoic acid

A 1 L jacketed three-necked round bottom flask equipped with a mechanical stirrer, rubber stopper with temperature probe, and a gas bubbler was filled with 100 ml of an ethanol solution containing 50.88 g of 7 . The solution was set to stir under nitrogen, diluted with 35 ml of ethanol and then 50 ml of 2-propanol and heated to about 60 ° C. 250 ml of water was then added to reach the cloudy point, and the cloudy solution was allowed to stand at about 60 ° C. for 75 minutes and then cooled to about 10 ° C. over about 1.5 hours. After 45 minutes, the mixture had two phases, which were diluted with 30 ml of additional 2-propanol. The mixture was stirred overnight at 10 ° C. under nitrogen and the resulting white fine suspension was filtered. The collected solid was washed with 100 ml of 9: 1 water: 2-propanol and dried in vacuo at about 50 ° C to 60 ° C to give 39.88 g of 7 as chalky white powder (78% recovery). The solid in the filtrate was filtered and dried to give 4.51 g of a pale yellow granular solid. HPLC indicated that this material was predominantly boronic acid.

6.11. Preparation of (S) -3- (4- (2-amino-6-chloropyrimidin-4-yl) phenyl) -2- (tert-butoxycarbonylamino) propanoic acid

The crude compound 7 (0.32 mol) was dissolved in ethanol (800 ml) and the resulting solution was concentrated to about 700 ml under reduced pressure and diluted with ethanol (1300 ml). To this solution 2-amino-4,6-dichloropyrimidine (74 g, 1.4 eq), bis (triphenylphosphine) palladium (II) dichloride (2.3 g, 1 mol%), and an aqueous potassium bicarbonate solution ( 97 g, 3 eq, 380 ml of water) was added. The mixture was heated at 75 ° C. to 80 ° C. for 2 hours, at which point HPLC analysis showed that the starting material was consumed completely. Ethanol was removed from the filtrate under reduced pressure to give an aqueous slurry (600 ml, additional water was added during distillation). The slurry was filtered and washed with 200 ml of water. The cake was dried under vacuum at 50 ° C. to give the recovered 2-amino-4,6-dichloropyrimidine as a tan solid (30 g, 41% of the original fill).

The filtrate was washed with ethyl acetate (400 ml) and diluted with 3: 1 THF / MTBE (600 ml). The mixture was acidified to pH about 3.5. The organic layer was washed with brine (300 ml) and concentrated to give crude product 8 as red oil (180 g). This oil was dissolved again in THF (300 ml), gloss-filtered and washed with THF (100 ml). The filtrate was diluted with isopropanol (400 ml) and the mixture was distilled to about 300 ml in air. Additional isopropanol (400 ml) was added and distillation continued until the volume reached about 500 ml. The mixture was then cooled to 45 ° C. over 1 hour, left for 2 hours and then cooled to room temperature over 1 hour. After further time, the slurry was filtered, washed with isopropanol (150 ml) and dried under vacuum at 50 ° C. to give product 8 as a pale pink solid (46.2 g, Boc-Tyr-OMe, yield from 4) . 37%). Purity by HPLC: 93.4%. Chiral purity:> 99% ee. Chiral analysis was performed on the corresponding methyl ester derivatives, which were prepared using trimethylsilyldiazomethane. Analytical pure samples were obtained by column chromatography (gradient 1:20 to 1:10 methanol / dichloromethane).

HPLC analysis of the mother liquor against a standard solution of compound 8 further showed 38 g of product 8 (30% yield from Boc-Tyr-OMe, 4 ). Product 8 was also partially recovered by further concentrating the mother liquor to give a total yield of 60% from Boc-Tyr-OMe, 4 .

6.12. Preparation of (S) -3- (4- (2-amino-6-chloropyrimidin-4-yl) phenyl) -2- (tert-butoxycarbonylamino) propanoic acid

A 22 L round-bottomed flask equipped with a mechanical stirrer, thermocouple attached to a temperature control, and a condenser with nitrogen line was added to compound 7 (850 g, 1 wt, 2.17 mol), 2-amino-4,6-dichloropyridine. Filled with midine (712.3 g, 2 equiv, right), and ethanol (13.6 L, 16 vol). The slurry was sparged with nitrogen for 10 minutes, then (Ph ₃ ) ₂ PdCl ₂ (18.3 g, 0.021 wt, strem) was added and the nitrogen sparging continued for 10 minutes. Subsequently, the reactor was charged with a solution of potassium bicarbonate (783 g, 3.6 equiv) in water (3.2 L, 3.7 vol), whereby gas evolution was observed. The mixture was heated to 75 ° C. for a total of 11.5 hours and then cooled to 45 ° C. overnight. HPLC analysis after 9.5 hours at 75 ° C. indicated that the residual amount of 7 (due to conversion) was about 3.0%. The reaction was cooled to 45 ° C. and stirred overnight, as a result of HPLC analysis showing that the residual amount of 7 was less than 1.0%.

The batch was then distilled over 15 hours at 45 ° C. under reduced pressure to yield 4 L to 5 L of a yellow slurry. The batch was then cooled overnight. Water was added (3 vol), heated to 45 ° C. and distillation continued for 1 hour until no more distillate was collected. The vacuum was released and water (3 vol) was added to the batch. After allowing to settle, the batch was filtered through a slurry of cellulose powder (20 μm, 0.2 wt.) In water (1 vol). Water (2 vol) was added to the remaining solids / slurry in the reactor, which was filtered through a sintered glass funnel. The filtrate was then further filtered through a cellulose pad to give 11.2 L of product solution (13.2 vol).

This solution was then transferred to a separatory funnel containing EtOAc (3.3 vol). After stirring and separation, the aqueous phase was transferred to a 22 L reactor and then charged with a solution of PBu ₃ (212 ml, 0.25 vol, 97%) in EtOAc (3.5 vol). The solution was heated at 50 ° C. for 2.5 h. Additional EtOAc (3.3 vol) was added to the reactor and the contents were charged to a separatory funnel and separated into two phases. The aqueous phase (41 ° C.) was again charged to a separatory funnel and washed with additional EtOAc (3.3 vol). After separating into two phases, the aqueous phase was charged to a 22 L reactor and heated to 45 ° C. Heptane (5 vol) was added to the reactor and the contents of the reactor were transferred to a separatory funnel and separated into two phases. The aqueous phase (11.2 L, 13.2 vol) was charged to a 22 L reactor, diluted to 14 vol with water and then a slurry of Dalco G-60 (0.2 wt) in water (1 vol) was charged to the reactor. The mixture was heated to 60 ° C. and stirred at 60 ° C. for 2 hours. Heat was blocked and the batch was stirred over the weekend. The batch was filtered through a Cellpure P300 (0.2 wt, Sigma) pad and washed with water (2 × 1.2 vol).

A 22 L round bottom flask equipped with a mechanical stirrer, thermocouple attached to a temperature controller, and a pH probe attached to a pH meter was filled with citric acid (127.5 g, 0.15 wt) and water (2 vol). The solution was heated to 40 ° C. and the pH of the solution was adjusted to 4.0 using sodium hydroxide 2 M solution. Citric acid solution (40 wt.%, 2 L) was charged to a further funnel and attached to the reactor. The basic solution of 8 was then transferred to the citric acid solution through the in-line filter of the peristaltic pump and the pH was maintained at pH 4.0 using 40% citric acid solution. Once the addition was complete, the batch was heated to 60 ° C. and stirred for 2 hours. The batch was then cooled overnight and the solid was filtered at 29 ° C. The cake was washed with water (2 × 2.5 vol) and dried at 45 ° C. to 50 ° C. for 24 hours to give 720 g of 8 (yield 84%) with a purity of 85.9% (AUC).

6.13. Purification of (S) -3- (4- (2-amino-6-chloropyrimidin-4-yl) phenyl) -2- (tert-butoxycarbonylamino) propanoic acid

When prepared as above, the title compound 8 typically contains about 6% dibasic acid impurity A and about 4% amination product B. While compound 8 can be used in the form of its crude product, it can also be purified using an approach as described below.

Method 1. Crude product 8 (10.0 g, 25.4 mmol, contains 90% purity, 6% A and 4% B ), i-PrOH / toluene (1: 1, 80 ml / 80 ml) in a 3-necked 250 ml RB flask , 8x / 8x) and tert-butylamine (13.4 ml, 5.0 equiv) were added. The resulting mixture was stirred, heated at 78 ° C. for 1 h, then slowly cooled to 0 ° C. and stirred for a further 1 h. The solid was collected by filtration and the cake washed with 20 ml of i-PrOH / toluene (1: 3). The cake was dried under vacuum to constant weight to afford the desired tert-butylamine salt of 8 as a pale yellow solid (8.8 g, yield 74%, purity 94%, A 3%, B 3%).

To a three-necked 250 ml RB flask, 8 tert-butylamine salts (20.0 g, 42.9 mmol) were added, followed by H ₂ O / THF / toluene (400 ml / 200 ml / 160 ml, 20x / 10x / 8x) Was added. The resulting mixture was heated to 60 ° C. and 6 M HCl was added slowly until the pH of the mixture reached 4.0. The mixture was cooled to room temperature and the organic layer was separated. The organic layer was washed with H ₂ O (100 ml, 5 ×), rotary evaporated and concentrated to a total volume of about 160 ml. The solid was collected by filtration and the cake washed with 20 ml of toluene. The cake was dried to constant weight under vacuum to give 8 as a pale yellow solid (15.0 g, yield 89%, purity 94%, A 3%, B 3%).

Method 2 . To a three-neck 250 ml RB flask add crude product 8 (containing 20.0 g, 42.9 mmol, 90% purity, 6% A and 4% B ), followed by THF / toluene (200 ml / 160 ml, 10 × / 8 ×) Was added. The resulting mixture was heated to 60 ° C. for 1 hour and cooled to room temperature. THF was removed by rotary evaporation to a total volume of about 160 ml. The solid was collected by filtration and the cake washed with 20 ml of toluene. The cake was dried under vacuum to constant weight to give 8 as a pale yellow solid (11.8 g, yield 70%, purity 92.8%, A 6.0%, B 1.3%).

To a three-necked 250 ml RB flask were added 8 (10.0 g, 25.4 mmol) and tert-butylamine (13.4 ml, 5 equiv), followed by i-PrOH / toluene (1: 1, 80 ml / 80 ml, 8x / 8x) was added. The resulting mixture was heated for 1 h until it became clear (78 ° C.), slowly cooled to 0 ° C. and stirred at 0 ° C. for an additional 1 hour. The solid was collected by filtration and the cake washed with 20 ml of i-PrOH / toluene (1: 3). The cake was dried under vacuum to constant weight to give 8 tert-butylamine salts as pale yellow solids (9.7 g, yield 82%, purity 96%, A 3.3%, B 0.6%).

To a three-necked 250 ml RB flask, 8 tert-butylamine salts (20.0 g, 42.9 mmol) were added, followed by H ₂ O / THF / toluene (400 ml / 200 ml / 160 ml, 20x / 10x / 8x) Was added. The resulting mixture was heated to 60 ° C. and 6 M HCl was added slowly until the pH of the mixture reached 4.0. The mixture was cooled to rt and the aqueous layer was separated. The organic layer was washed with H ₂ O (100 ml, 5 ×) and concentrated to about 160 ml total volume by rotary evaporation. The solid was collected by filtration and the cake washed with 20 ml of toluene. The cake was dried to constant weight under vacuum to afford 8 as a pale yellow solid (15 g, yield 88%, purity 96%, A 3.3%, B 0.5%).

Method 3. To a three-necked 3 L RB flask was added an aqueous potassium salt solution containing about 50 g of 8 (90%, A 6%, B 4%, all normalized AUC), followed by THF / toluene (500 ml / 400 ml, 10x / 8x) was added. The resulting mixture was heated to 60 ° C. and 6 M HCl was added slowly until the pH of the mixture reached 4.0. The mixture was cooled to rt and the aqueous layer was separated. The organic layer was washed with H ₂ O (250 ml, 5 ×) and concentrated by rotary evaporation to about 400 ml in total volume to give a slurry of 8 in about 8 × toluene.

To a three-neck 3 L RB flask was added slurry (400 ml in 8x toluene) and tert-butylamine (67 ml, 5.0 equiv) followed by i-PrOH (400 ml, 8x). The resulting mixture was heated at 78 ° C. for 1 hour, cooled to 0 ° C. and stirred at 0 ° C. for an additional hour. The solid was collected by filtration and the cake washed with 100 ml of i-PrOH / toluene (1: 3). The cake was dried under vacuum to constant weight to give 8 tert-butylamine salts as pale yellow solids (42.4 g, yield 72%, purity 95%, A 3.2%, B 1.9%).

To a three-necked 250 ml RB flask were added 8 tert-butylamine salts (42.4 g, 91.0 mmol), followed by H ₂ O / THF / toluene (1000 ml / 500 ml / 400 ml, 20 × / 10 × / 8 ×) Was added. The resulting mixture was heated to 60 ° C. and 6 M HCl was added slowly until the pH reached 4.0. The mixture was cooled to room temperature. The organic layer was separated and washed with H ₂ O (250 ml, 5 ×). The organic solution was concentrated by rotary evaporation to a total volume of about 400 ml. The solid was collected by filtration and the cake washed with 100 ml of toluene. The cake was dried to constant weight in vacuo to afford 8 as a pale yellow solid (35.4 g, yield 89.5%, purity 96%, A 2.9%, B 1.6%).

Method 4. 8 (198.6 mg, 0.5 mmol) and cicononidine (167.1 mg) were added to the test tube followed by acetonitrile (7.5 ml). The resulting mixture was heated to clear, cooled to rt and stirred for a further 2 h. The solid was collected by filtration and the cake washed with 1 ml of MTBE. The cake was dried under vacuum to constant weight to give the final product (208 mg, yield 68%, purity 92%, A 4.4%, B 1.4%).

6.14. Of (S) -3- (4- (2-amino-6-chloropyrimidin-4-yl) phenyl) -2- (tert-butoxycarbonylamino) propanoic acid (8) with potassium carbonate as the base Produce

500 ml 3-neck round bottom flask equipped with mechanical stirrer and temperature control 2-amino-4,6-dichloropyrimidine (12.57 g, 1.5 equiv), boronate compound 7 (20.00 g, 51.1 mmol) , Potassium carbonate (21.19 g, 3.0 equiv) and ethanol / water (200 ml, 5: 1 by volume) were charged. The mixture was stirred and catalyst bis (triphenylphosphine) palladium (II) dichloride (359 mg, 1 mol%) was added. The mixture was heated to 80 ° C. and stirred for 2 hours. The reaction was cooled to rt and diluted with water (100 ml). The mixture was then concentrated under reduced pressure to remove most of the ethanol and 1 N NaOH (60 ml) was added. The mixture was extracted twice with ethyl acetate (2 × 200 ml) and the aqueous layer was acidified to pH about 3 with 1 N HCl. The mixture was extracted twice with ethyl acetate (200 ml and 100 ml respectively), the combined organic layers were concentrated and the residue was purified by column chromatography (gradient 1:20 to 1:10 methanol / dichloromethane), compound 8 was obtained as a pale yellow solid (15.92 g, 79%).

6.15. (S) -2- (tert-butoxycarbonylamino) -3- (4- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) phenyl) Preparation of (S) -3- (4- (2-amino-6-chloropyrimidin-4-yl) phenyl) -2- (tert-butoxycarbonylamino) propanoic acid with lithium salt of propanoic acid

During the preparation of compound 7 , the isolation of the free acid can optionally be omitted. Thus, an aqueous lithium salt solution of compound 7 in 100 ml of water prepared from 5.0 g of Boc-Tyr-OMe ( 4 , 17 mmol) was added with 2-amino-4,6-dichloropyrimidine (3.3 g, 1.2 eq), potassium bicarbonate. (5.0 g, 3 eq), bis (triphenylphosphine) palladium (II) dichloride (60 mg, 0.5 mol%), and 100 ml of ethanol. The resulting mixture was heated at 70 ° C. for 5 hours. Additional 2-amino-4,6-dichloropyrimidine (1.1 g, 0.4 eq) was added and heating continued at 70 ° C. for a further 2 hours. HPLC analysis showed about 94% conversion. After cooling and filtration, the filtrate was analyzed by HPLC against a standard solution of compound 8 . The analysis showed that 3.9 g of compound 8 was contained in the solution (yield 59% from compound 4 ).

6.16. For the preparation of (S) -3- (4- (2-amino-6-chloropyrimidin-4-yl) phenyl) -2- (tert-butoxycarbonylamino) propanoic acid using potassium carbonate as base Alternative course

Boronic acid compound 11 (Ryscor Science, Inc., North Carolina, USA, 1.0 g, 4.8 mmol) and potassium carbonate (1.32 g, 2 eq) were added to aqueous ethanol (15 ml of ethanol and water 8 ml). Di-tert-butyldicarbonate (1.25 g, 1.2 eq) was added in one portion. HPLC analysis after 30 min stirring at room temperature showed that starting compound 11 was consumed completely. 2-amino-4,6-dichloropyrimidine (1.18 g, 1.5 eq) and catalyst bis (triphenylphosphine) palladium (II) dichloride (34 mg, 1 mol%) were added and the resulting mixture was 65 Heated at C-70 C for 3 h. HPLC analysis showed that Compound 12 was consumed completely. After concentration and filtration, HPLC analysis of the resulting aqueous solution of the standard solution of compound 8 showed that the compound 8 1.26 g (yield 67%).

6.17. Of (S) -3- (4- (2-amino-6-chloropyrimidin-4-yl) phenyl) -2- (tert-butoxycarbonylamino) propanoic acid using potassium carbonate / potassium bicarbonate as the base Alternative Process for Manufacturing

Boronic acid compound 11 (10 g, 48 mmol) and potassium bicarbonate (14.4 g, 3 eq) were mixed in aqueous ethanol (250 ml of ethanol and 50 ml of water). Di-tert-butyldicarbonate (12.5 g, 1.2 eq) was added in one portion. HPLC analysis showed that the reaction was not complete even after stirring overnight at room temperature. Potassium carbonate (6.6 g, 1.0 eq) and additional di-tert-butyldicarbonate (3.1 g, 0.3 eq) were added. HPLC analysis after 2.5 h stirring at room temperature showed the starting compound 11 was consumed completely. 2-amino-4,6-dichloropyrimidine (11.8 g, 1.5 eq) and catalyst bis (triphenylphosphine) -palladium (II) dichloride (0.34 g, 1 mol%) were added and the resulting mixture was Heated at 75 ° C. to 80 ° C. for 2 hours. HPLC analysis showed that Compound 12 was consumed completely. The mixture was concentrated under reduced pressure and filtered. The filtrate was washed with ethyl acetate (200 ml) and diluted with 3: 1 THF / MTBE (120 ml). This mixture was acidified to pH about 2.4 with 6 N hydrochloric acid. The organic layer was washed with brine and concentrated under reduced pressure. The residue was precipitated in isopropanol, filtered and dried under vacuum at 50 ° C. to give compound 8 as an off-white solid (9.0 g, 48% yield). Purity by HPLC analysis: 92.9%. The mother liquor was concentrated to give 2.2 g more off-white powder (yield 12%). Purity by HPLC analysis: 93.6%.

6.18. (S) -3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3'-methoxybiphenyl-4-yl) ethoxy) pyri) Preparation of midin-4-yl) phenyl) -2- (tert-butoxycarbonylamino) propanoic acid

In a 250 ml three-necked round bottom flask equipped with a mechanical stirrer and temperature control, monochloride 8 (20.39 g, 51.9 mmol), alcohol 3 (17.58 g, 1.2 equiv), cesium carbonate (84.55, 5.0 equiv) and Dioxane (205 ml) was charged. The mixture was heated to 100 ° C. and stirred for 17 h. The reaction was cooled to rt and diluted with water (80 ml). The organic layer was collected, diluted with two phases, diluted with ethyl acetate (200 ml) and washed with a mixture of brine (50 ml) and 1N HCl (50 ml). The organic layer was concentrated and the residue was purified by column chromatography (gradient: 1:30 to 1:20 methanol / dichloromethane and 0.5% acetic acid) to give compound 9 as a yellow solid. This solid was recrystallized from EtOH and heptane to give 21.78 g of a pale yellow solid. The mother liquor was further crystallized to give 2.00 g of a pale yellow solid (23.78 g total, 72% yield). Chiral analysis of the corresponding methyl ester derivative prepared using trimethylsilyldiazomethane showed no detectable amount of diastereomers.

6.19. (S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3'-methoxybiphenyl-4-yl) Preparation of ethoxy) pyrimidin-4-yl) phenyl) propanoic acid

To a 500 ml round bottom flask was added compound 9 (20.00 g, 31.32 mmol) and THF (100 ml). The solid was dissolved with stirring and 6 N hydrochloric acid (100 ml) was added slowly. The mixture was then stirred at rt for 14 h. The reaction was diluted with water (100 ml) and most of THF was removed under reduced pressure. The resulting aqueous solution was then transferred to a 500 ml three-necked round bottom flask equipped with a mechanical stirrer, pH meter, temperature controller and addition funnel. 50% aqueous sodium hydroxide solution was added slowly at 60 ° C. until pH = 4, then 1 N aqueous sodium hydroxide solution was added until pH reached 6.5. The mixture was stirred for an additional 30 minutes at 60 ° C., the solids were collected by filtration and oven dried under vacuum to afford compound 10 (16.30 g, yield 96%) as a pale yellow solid.

6.20. (S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3'-methoxybiphenyl-4-yl) One-pot preparation of ethoxy) pyrimidin-4-yl) phenyl) propanoic acid

In a 3-necked 250 ml round bottom flask equipped with a mechanical stirrer and temperature control compound 3 (8.62 g, 1.2 equiv), 8 (10.00 g, 25.46 mmol), tetrabutylammonium bisulfate (0.86 g, 10 mol) %), And cesium carbonate (29.04 g, 3.5 equiv). Dioxane (50 ml) was added and the resulting mixture was heated at 100 ° C. for 18 hours. HPLC analysis showed 99% conversion of starting material 8 . The mixture was cooled to 60 ° C. and water (60 ml) was added. The upper organic layer was diluted with THF (80 ml), washed with brine (50 ml), transferred to a 500 ml round bottom flask and 80 ml of 6 N hydrochloric acid was added. The mixture was stirred at rt for 16 h. LC-MS analysis of the reaction mixture showed that intermediate compound 9 was consumed completely. The reaction mixture was transferred to a 500 ml separatory funnel. The round bottom flask was washed with water (2 × 40 ml) and the wash was transferred to a funnel. The mixture was washed with ethyl acetate (2 × 100 ml), the aqueous layer collected and concentrated under vacuum at 80 mbar at 40 ° C. (bath temperature) to remove any residual organic solvent. The resulting aqueous solution was then transferred to a 500 ml three-necked round bottom flask equipped with a mechanical stirrer, pH meter, temperature controller and addition funnel. At 60 ° C., 50% aqueous sodium hydroxide solution was added slowly to pH = 4, then 1 N aqueous sodium hydroxide solution was added until the pH reached 6.5. The mixture was stirred for an additional 30 minutes at 60 ° C. and the yellow solid was collected by filtration. HPLC analysis of this solid showed that the purity was about 95%. The solid was dried overnight under vacuum at 50 ° C. to afford crude 10 as a yellow solid (9.48 g, 69% total yield).

The solid (9.48 g) was transferred to a 500 ml round bottom flask and water (95 ml) was added. The mixture was heated at 80 ° C. (bath temperature) and THF (40 ml) was added to dissolve the solids. Subsequently, most of the THF was removed at 80 ° C. under vacuum. Acetonitrile (80 ml) was added to the precipitate, stirred at 80 ° C. for 2 hours, cooled to room temperature and then stirred at 0 ° C. for 30 minutes. The solid was collected by filtration and washed with water (2 x 50 ml) to afford compound 10 as a pale yellow solid (8.53 g, 90% recovery, 62% total yield). HPLC analysis showed that the purity exceeded 99%.

6.21. (S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3'-methoxybiphenyl-4-yl) One-pot preparation of ethoxy) pyrimidin-4-yl) phenyl) propanoic acid

Fill a 22 L round bottom flask with 1,4-dioxane (4 vol) with a mechanical stirrer, thermocouple attached to a temperature control, and a condenser with nitrogen line, followed by Cs ₂ CO ₃ (2.03 kg, 3.5 equiv), compound 3 (603 g, 1.2 equiv) and tetrabutylammonium bisulfate (102.8 g, 0.147 wt) were added. The slurry was slowly heated to 70 ° C., and then a slurry of compound 8 (700.0 g, 1.782 mol, 1 wt) in 1,4-dioxane (1.5 vol) was added in portions over 10 minutes. The beaker containing 8 was washed with 1,4-dioxane (0.5 vol) and added to the reactor. The reaction started to thicken briefly after stirring for 15-30 minutes, but the entire batch was stirrable. The control was heated at 78 ° C. overnight, then at 98 ° C. for 8 hours and then at 85 ° C. overnight. HPLC analysis showed that 8 remained 2.1%. The reaction was quenched with water (6 vol) at 78 ° C. and then further cooled. At 42 ° C. the batch was transferred to a separating funnel and separated into two phases. The organic phase was then diluted with THF (8 vol) and washed with brine (5 vol). The phases were separated and the organic phase was washed with brine (5 vol). The phases were separated and the organic phase (9.5 L) was transferred to a 22 L reactor. 6 N HCl solution (11.4 vol) was added and the batch was heated at 40 ° C. to 45 ° C. for 2 hours. HPLC analysis showed the reaction was complete and Dalco G-60 (0.33 wt.) And water (2 vol) were added. The batch was stirred at 40 ° C. over the weekend and then heated to 60 ° C. The reaction mixture was filtered through a PTFE cloth at 60 ° C. and the reactor was washed with water (6 vol). The wash was heated to 60 ° C. and washed through a dalco pad. The filtrate was then passed through a 0.3 μm in-line filter and washed twice with IPAc (10 vol, 8.8 vol). The aqueous phase was then concentrated using a 20 L rotary evaporator at 45 ° C. under reduced pressure until the mixture became cloudy (2-3 hours). The volume of collected distillate was approximately 3.3 L. The batch was then returned back to the 22 L reactor and left at 40 ° C. overnight.

The batch was heated to 60 ° C. whereby the batch changed from cloudy to clear. A separate 22 L reactor was charged with water (1.6 vol) and 85% phosphoric acid (0.24 vol) and the pH was adjusted to 6.5 using 50% NaOH solution (approximately 0.3 vol). The acidic product solution was then transferred via a peristaltic pump to a reactor containing a buffer solution of pH 6.5 and 50% NaOH was added (approximately 3.5 vol) to maintain the pH within 6 and 7. The temperature of the reactor was maintained at 55 ° C. to 65 ° C. (additional 2 hours). Once the addition was complete, the slurry was heated at 60 ° C. to 65 ° C. for 90 minutes, filtered and washed with water (2 × 6.7 vol). The wet cake was dried in a vacuum oven at 55 ° C. for 39 hours to give 635 g of crude product 10 as a yellow solid (yield 66%). The purity of the product was 93.2% (AUC).

6.22. (S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3'-methoxybiphenyl-4-yl) Purification of ethoxy) pyrimidin-4-yl) phenyl) propanoic acid

A 22 L round bottom flask equipped with a mechanical stirrer, thermocouple attached to a temperature control, and a condenser with nitrogen line was charged with crude product 10 (630 g) followed by the addition of THF (5 vol). The slurry was heated to 65 ° C. After 30 minutes a 5 N to 6 N HCl solution (0.47 L, 0.746 vol) in IPA was added and the solids were slowly dissolved. The orange solution was heated at 65 ° C. for 30 minutes and IPA (10 vol) was added slowly while maintaining the temperature at 60 ° C. to 70 ° C. When the addition was complete, the mixture was stirred for 20 minutes, then IPAc (10 vol) was added slowly while maintaining the temperature at 60 ° C to 70 ° C. Once the addition was complete, the thick slurry was stirred at 65 ° C. for 1 hour and then cooled to 27 ° C. over 4.5 hours. The solid was filtered off and washed with IPA (2 × 3 vol). The product was dried in a vacuum oven at 55 ° C. for 15 hours to give 630 g of diHCl salt 10 with a purity of 95.0% (AUC) (yield 88%).

A 12 L round bottom flask equipped with a mechanical stirrer, thermocouple attached to a temperature control, and a pH probe attached to a pH meter was added to diHCl salt 10 (620 g, 1 wt) followed by 1 M aqueous NaOH solution (10 vol). Filled. The mixture was heated to 40 ° C., stirred until all solids dissolved (2 hours) and then transferred to a 10 L carboy. The 12 L round bottom flask was washed with water and then 85% phosphoric acid (124 ml, 0.2 vol) and water (1.3 vol) were charged to the reactor. The pH was adjusted to 6.5 with 50% NaOH (0.24 vol) and then heated to 65 ° C. The product solution in the carboy was transferred to a pH buffer solution via a peristaltic pump and the pH was maintained at 6-7 by adding 6 M HCl aqueous solution (0.67 L). Once the addition was complete, the slurry was heated at 65 ° C. for 3 hours and the solids were filtered off. The cake was washed with water (3 × 5 vol) and then dried in a vacuum oven at 55 ° C. for 41 hours to give 473 g of 10 as a light yellow solid with a purity of 97.7% (AUC) (yield 87%).

6.23. Crystalline (S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3'-methoxybiphenyl-4-yl Preparation of) ethoxy) pyrimidin-4-yl) phenyl) propanoate tosylate anhydride

(S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3'-methoxybiphenyl-4-yl) Ethoxy) pyrimidin-4-yl) phenyl) propanoate free base (11.8 g assay, 22.0 mmol) was added TsOH.H ₂ O (4.60 g, in THF (35 ml) and water (2.8 ml) at 40 ° C. 24.0 mmol) in solution. Acetonitrile (35 ml) was added and the mixture was aged until a slurry was obtained. Additional acetonitrile (105 ml) was added slowly over 1 hour. The mixture was aged at 40 ° C. for 2 hours and then slowly cooled to 20 ° C. over 3 hours. After maturing for 5 hours at 20 ° C., the mixture was filtered and washed with ACN / THF / H ₂ O (50/10/1 ml). The filter cake was swept slowly with nitrogen at 40 ° C. in a vacuum oven to dryness to afford the title compound.

6.24. Crystalline (S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3'-methoxybiphenyl-4-yl Preparation of (ethoxy) pyrimidin-4-yl) phenyl) propanoate tosylate monohydrate

500 ml of 3-sphere RBF were added to (S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3 ') at room temperature. -Methoxybiphenyl-4-yl) ethoxy) pyrimidin-4-yl) phenyl) propanoic acid (zwitterion, 10 g, 18.6 mmol), p-toluenesulfonic acid monohydrate (3.94 g, 20.4 mmol), Filled with CH ₃ CN (50 ml, 5 ×) and water (10 ml, 1 ×). The suspension was heated to 75 ° C. under gentle reflux. AcOH was then added (20 ml, 2 ×) to give a clear yellow solution (the first 10 ml of AcOH already cleared the suspension). To this clear solution was added CH ₃ CN (60 ml, 6 ×), and the solution was turbid at the time of seeding, followed by additional CH ₃ CN (40 ml, 4 ×). The resulting slurry was stirred at 80 ° C. for 40 minutes, then it was cooled to room temperature with stirring, stirred overnight, filtered and washed with CH ₃ CN / water (15/1, 40 ml, 4 ×). The wet pale yellow cake was dried under vacuum at 40 ° C. overnight to yield an off-white solid (9.3 g, recovery 68.6%, purity 98.8% by the 40 minute release method, KF = 0.93%). When covered with a paper cover and left on a laboratory bench over four days, KF increased to 3.262% and weight increased to 9.61 g (71%).

6.25. Crystalline (S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3'-methoxybiphenyl-4-yl Preparation of (ethoxy) pyrimidin-4-yl) phenyl) propanoate tosylate dihydrate

(S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3'-methoxybiphenyl-4-yl) Ethoxy) pyrimidin-4-yl) phenyl) propanoate (120.0 g, 88 w%, 105.6 g active, 196 mmol) was added to TsOH.H ₂ O in a mixture of THF (240 ml) and water (48 ml). (39.8 g, 209 mmol) was added to the solution. The mixture was heated to 50 ° C. to obtain a homogeneous solution. Approximately 120 ml of a mixture of ACN / water (1200/60 ml) was added and the mixture was seeded with the title compound (0.63 g). After aging at 40 ° C. for 1 hour, a good slurry was obtained. The remaining ACN / water mixture was added slowly at 40 ° C. over 3 hours, the slurry was aged at 40 ° C. for 2 hours, then slowly cooled to 20 ° C. and aged overnight. The solid was collected by filtration and the filter cake washed with 5/1 ACN / THF containing about 5 vol% (500 ml) of water. Air drying at room temperature overnight gave 138.5 g of the product as a white solid (99.5 A%, 93.4% yield corrected for purity). 6.5% was lost in the mother liquor and wash liquor. KF of the solid was 4.4%.

6.26. Crystalline (S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3'-methoxybiphenyl-4-yl Preparation of (ethoxy) pyrimidin-4-yl) phenyl) propanoate maleate

(S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3'-methoxybiphenyl-) in 1 ml of methanol To the solution of 4-yl) ethoxy) pyrimidin-4-yl) phenyl) propanoate (13.3 mg) was added maleic acid (3.2 mg). The mixture was heated under gentle reflux and then cooled to room temperature to afford the title compound.

All references mentioned above (eg patents and patent applications) are hereby incorporated by reference in their entirety.

Claims (31)

(S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3'-methoxybiphenyl-4-yl) Crystalline compound that is ethoxy) pyrimidin-4-yl) phenyl) propanoic acid or a salt thereof. (S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3'-methoxybiphenyl-4-yl) Crystalline compound that is ethoxy) pyrimidin-4-yl) phenyl) propanoate tosylate. The crystalline compound of claim 2, which is an anhydride. The crystalline compound of claim 3, wherein the melting point is about 241 ° C. 5. The crystalline compound of claim 3, having an X-ray powder diffraction pattern comprising peaks at one or more of 2θ angles of about 3.5, 7.0, 8.6, 10.9, 13.5, 14.0, 15.1, 17.3, and / or 20.5. The compound of claim 3 having an X-ray powder diffraction pattern substantially the same as that shown in FIG. 1. The crystalline compound of claim 2, which is a hydrate. 8. The crystalline compound of claim 7, which is a monohydrate. The crystalline compound of claim 8, wherein the melting point is about 221 ° C. 10. The compound of claim 8, having an X-ray powder diffraction pattern comprising peaks at one or more of 2θ angles of about 3.6, 8.2, 8.7, 13.1, 14.5, 17.5, 18.0, 19.9, and / or 21.4. The compound of claim 8, having an X-ray powder diffraction pattern substantially the same as that shown in FIG. 2. The compound of claim 8, having a Raman spectrum substantially the same as that shown in FIG. 3. 8. The crystalline compound of claim 7, which is a dihydrate. The crystalline compound of claim 13, wherein the melting point is about 238 ° C. 15. The crystalline compound of claim 13, having an X-ray powder diffraction pattern comprising peaks at one or more of the 2θ angles of about 8.6, 9.0, 17.2, 17.8, 18.6, 21.6, 25.2, and / or 26.9. The compound of claim 13, having an X-ray powder diffraction pattern substantially the same as that shown in FIG. 4. (S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3'-methoxybiphenyl-4-yl) Crystalline compound, which is ethoxy) pyrimidin-4-yl) phenyl) propanoate maleate. A pharmaceutical dosage form comprising the crystalline compound of claim 1. The compound of claim 18, wherein the crystalline compound is (S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3'-). Pharmaceutical dosage form, which is methoxybiphenyl-4-yl) ethoxy) pyrimidin-4-yl) phenyl) propanoate tosylate. The compound of claim 18, wherein the crystalline compound is (S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3'-). Pharmaceutical dosage form, which is methoxybiphenyl-4-yl) ethoxy) pyrimidin-4-yl) phenyl) propanoate maleate. (S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3'-methoxybiphenyl-4-yl) A solution containing ethoxy) pyrimidin-4-yl) phenyl) propanoic acid and a pharmaceutically acceptable acid is heated to give (S) -2-amino-3- (4- (2-amino-6-(( R) -2,2,2-trifluoro-1- (3'-methoxybiphenyl-4-yl) ethoxy) pyrimidin-4-yl) phenyl) propanoic acid;
The solubility of the salt in the solution is (S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3'-methoxy) Reducing under conditions sufficient to provide a crystalline salt of biphenyl-4-yl) ethoxy) pyrimidin-4-yl) phenyl) propanoic acid; And
Isolating Crystalline Salts
(S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3'-methoxybiphenyl-) A process for preparing the crystal salt of 4-yl) ethoxy) pyrimidin-4-yl) phenyl) propanoic acid. The method of claim 21, wherein the solution comprises THF and water. The method of claim 21, wherein the pharmaceutically acceptable acid is p-toluenesulfonic acid or maleic acid. The method of claim 21, wherein the solubility of the salt is reduced by adding antisolvent and cooling the solution. The method of claim 24, wherein the antisolvent is acetonitrile. Water, (S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3'-methoxybiphenyl-4- 1) heating a solution comprising ethoxy) pyrimidin-4-yl) phenyl) propanoic acid and p-toluenesulfonic acid monohydrate;
Adding an antisolvent to the solution to provide a mixture;
Cooling the mixture; And
Crystalline (S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3'-methoxybiphenyl-4) -Yl) ethoxy) pyrimidin-4-yl) phenyl) propanoic acid tosylate
Crystal (S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2-trifluoro-1- (3'-methoxybiphenyl) containing 4-yl) ethoxy) pyrimidin-4-yl) phenyl) propanoic acid tosylate. The method of claim 26, wherein the antisolvent is acetonitrile. Determination in patients in need of treatment, prevention or management of diseases or disorders mediated by peripheral serotonin (S) -2-amino-3- (4- (2-amino-6-((R) -2,2,2) Administering a therapeutically or prophylactically effective amount of -trifluoro-1- (3'-methoxybiphenyl-4-yl) ethoxy) pyrimidin-4-yl) phenyl) propanoic acid or salts thereof, A method of treating, preventing or managing a disease or disorder mediated by peripheral serotonin. The method of claim 28, wherein the disease or disorder is carcinoid syndrome. The method of claim 28, wherein the disease or disorder is a gastrointestinal disease or disorder. 31. The method of claim 30, wherein the disease or disorder is irritable bowel syndrome.

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